Arg-gly-asp-mannose-6-phosphate inhibits activation and proliferation of hepatic stellate cells in v

来源 :World Journal of Gastroenterology | 被引量 : 0次 | 上传用户:mythology_leonie
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AIM:To investigate the effect of arg-gly-asp-mannose-6phosphate(RGD-M6P)on the activation and proliferationof primary hepatic stellate cells in vitro.METHODS:Hepatic stellate cells(HSCs)were isolatedfrom rats by in situ collagenase perfusion of liver and18% Nycodenz gradient centrifugation and cultured onuncoated plastic plates for 24 h with DMEM containing10% fetal bovine serum(FBS/DMEM)before the culturemedium was substituted with 2% FBS/DMEM for another24 h.Then,HSCs were cultured in 2% FBS/DMEM withtransforming growth factor 131,M6P,RGD,or RGD-M6P,respectively.Cell morphology was observed underinverted microscope,smooth muscle α-actin(α-SMA)was detected by immunocytochemistry,type Ⅲprocollagen(PCⅢ)in supernatant was determined byradioimmunoassay,and the proliferation rate of HSCswas assessed by flow cytometry.RESULTS:RGD-M6P significantly inhibited themorphological transformation and the α-SMA and PCⅢ expressions of HSCs in vitro and also dramaticallyprevented the proliferation of HSCs in vitro.Such effectswere remarkably different from those of RGD or M6P.CONCLUSION:The new compound,RGD-M6P,whichhas a dramatic effect on primary cultured HSCs invitro,can inhibit the transformation of HSCs in culturecaused by TGFβ1,suppresses the expression of PCⅢand decreases proliferation rate of HSC.RGD-M6P canbe applied as a selective drug carrier targeting at HSCs,which may be a new approach to the prevention andtreatment of liver fibrosis. AIM: To investigate the effect of arg-gly-asp-mannose-6 phosphate (RGD-M6P) on the activation and proliferation of primary hepatic stellate cells in vitro. METHODS: Hepatic stellate cells (HSCs) were isolatedfrom rats by in situ collagenase perfusion of liver and 18% Nycodenz gradient centrifugation and cultured onuncoated plastic plates for 24 h with DMEM containing 10% fetal bovine serum (FBS / DMEM) before the culture medium was substituted with 2% FBS / DMEM for another 24 h. / DMEM with transforming growth factor 131, M6P, RGD, or RGD-M6P, respectively. Cell morphology was observed under inverted microscope, smooth muscle α-actin (α-SMA) was detected by immunocytochemistry, type Ⅲ protocollagen (PC Ⅲ) in supernatant was determined by radioimmunoassay , and the proliferation rate of HSCs were estimated by flow cytometry. RESULTS: RGD-M6 significantly inhibited themorphological transformation and the α-SMA and PCⅢ expressions of HSCs in vitro and also dramaticallyprevented the prolifer of HSCs in culture of HSCs in vitro remarkably different from those of RGD or M6P.CONCLUSION: The new compound, RGD-M6P, which has a dramatic effect on primary cultured HSCs invitro, can inhibit the transformation of HSCs in culturecaused by TGFβ1, suppresses the expression of PCⅢand decreasing proliferation rate of HSC.RGD-M6P canbe applied as a selective drug carrier targeting at HSCs, which may be a new approach to the prevention and treatment of liver fibrosis.
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